Bottom Line:
A comparison of transcriptional profiles from Müller cells at one or three days after CNTF treatment showed an increase in the number of transcribed genes as well as a change in the expression pattern.Further analysis of gene profiles revealed 20-30% overlap in the transcription pattern among Müller cells, astrocytes and the RPE.Our studies provide novel molecular insights into biological functions of Müller glial cells in mediating cytokine response.

Affiliation: Department of Ophthalmology, Northwestern University Feinberg Medical School, Chicago, Illinois, United States of America.

ABSTRACT

Background: Ciliary neurotrophic factor (CNTF), a member of the interleukin-6 cytokine family, has been implicated in the development, differentiation and survival of retinal neurons. The mechanisms of CNTF action as well as its cellular targets in the retina are poorly understood. It has been postulated that some of the biological effects of CNTF are mediated through its action via retinal glial cells; however, molecular changes in retinal glia induced by CNTF have not been elucidated. We have, therefore, examined gene expression dynamics of purified Müller (glial) cells exposed to CNTF in vivo.

Methodology/principal findings: Müller cells were flow-sorted from mgfap-egfp transgenic mice one or three days after intravitreal injection of CNTF. Microarray analysis using RNA from purified Müller cells showed differential expression of almost 1,000 transcripts with two- to seventeen-fold change in response to CNTF. A comparison of transcriptional profiles from Müller cells at one or three days after CNTF treatment showed an increase in the number of transcribed genes as well as a change in the expression pattern. Ingenuity Pathway Analysis showed that the differentially regulated genes belong to distinct functional types such as cytokines, growth factors, G-protein coupled receptors, transporters and ion channels. Interestingly, many genes induced by CNTF were also highly expressed in reactive Müller cells from mice with inherited or experimentally induced retinal degeneration. Further analysis of gene profiles revealed 20-30% overlap in the transcription pattern among Müller cells, astrocytes and the RPE.

Conclusions/significance: Our studies provide novel molecular insights into biological functions of Müller glial cells in mediating cytokine response. We suggest that CNTF remodels the gene expression profile of Müller cells leading to induction of networks associated with transcription, cell cycle regulation and inflammatory response. CNTF also appears to function as an inducer of gliosis in the retina.

pone-0020326-g007: Gene networks common to Müller cells, astrocytes and RPE.(a) Most significant canonical pathways; (b) Most significant biological functions for the same list of genes; and (c) Genes in this network are responsible mitochondrial function and metabolism.

Mentions:
IPA of the 381 genes (common to Müller cells, astrocytes and RPE) showed that the top canonical pathways were concerned with oxidative phosphorylation, mitochondrial function, glycolysis/gluconeogenesis, hypoxia signaling and ubiquinone biosynthesis. This finding is in accord with the known supportive roles of the three cell types in energy metabolism in the retina [41], [57]. The top biological functions involve protein synthesis, RNA post-transcriptional modification, and energy production (Fig. 7).

pone-0020326-g007: Gene networks common to Müller cells, astrocytes and RPE.(a) Most significant canonical pathways; (b) Most significant biological functions for the same list of genes; and (c) Genes in this network are responsible mitochondrial function and metabolism.

Mentions:
IPA of the 381 genes (common to Müller cells, astrocytes and RPE) showed that the top canonical pathways were concerned with oxidative phosphorylation, mitochondrial function, glycolysis/gluconeogenesis, hypoxia signaling and ubiquinone biosynthesis. This finding is in accord with the known supportive roles of the three cell types in energy metabolism in the retina [41], [57]. The top biological functions involve protein synthesis, RNA post-transcriptional modification, and energy production (Fig. 7).

Bottom Line:
A comparison of transcriptional profiles from Müller cells at one or three days after CNTF treatment showed an increase in the number of transcribed genes as well as a change in the expression pattern.Further analysis of gene profiles revealed 20-30% overlap in the transcription pattern among Müller cells, astrocytes and the RPE.Our studies provide novel molecular insights into biological functions of Müller glial cells in mediating cytokine response.

Affiliation:
Department of Ophthalmology, Northwestern University Feinberg Medical School, Chicago, Illinois, United States of America.

ABSTRACT

Background: Ciliary neurotrophic factor (CNTF), a member of the interleukin-6 cytokine family, has been implicated in the development, differentiation and survival of retinal neurons. The mechanisms of CNTF action as well as its cellular targets in the retina are poorly understood. It has been postulated that some of the biological effects of CNTF are mediated through its action via retinal glial cells; however, molecular changes in retinal glia induced by CNTF have not been elucidated. We have, therefore, examined gene expression dynamics of purified Müller (glial) cells exposed to CNTF in vivo.

Methodology/principal findings: Müller cells were flow-sorted from mgfap-egfp transgenic mice one or three days after intravitreal injection of CNTF. Microarray analysis using RNA from purified Müller cells showed differential expression of almost 1,000 transcripts with two- to seventeen-fold change in response to CNTF. A comparison of transcriptional profiles from Müller cells at one or three days after CNTF treatment showed an increase in the number of transcribed genes as well as a change in the expression pattern. Ingenuity Pathway Analysis showed that the differentially regulated genes belong to distinct functional types such as cytokines, growth factors, G-protein coupled receptors, transporters and ion channels. Interestingly, many genes induced by CNTF were also highly expressed in reactive Müller cells from mice with inherited or experimentally induced retinal degeneration. Further analysis of gene profiles revealed 20-30% overlap in the transcription pattern among Müller cells, astrocytes and the RPE.

Conclusions/significance: Our studies provide novel molecular insights into biological functions of Müller glial cells in mediating cytokine response. We suggest that CNTF remodels the gene expression profile of Müller cells leading to induction of networks associated with transcription, cell cycle regulation and inflammatory response. CNTF also appears to function as an inducer of gliosis in the retina.